December 30, 2014 Archives • 2014 • vol.3 •197-202 PROTECTIVE EFFECT OF HERACLEUM PERSICUM ALCOHOLIC EXTRACT ON CYCLOPHOSPHAMIDE-INDUCED GAMETOGENIC DAMAGE IN RATS F. Dalouchi1*, S. M. Banan Khojasteh1, G. Dehghan1, D. Mohhamadnezhad2, S. Rostampour1, N. Farsad3 1Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran 2 Faculty of medicine, Medical Sciences, University of Tabriz, Tabriz, Iran 3 Department of Botany, Faculty of Science, Tabriz University, Tabriz, Iran *[email protected] Abstract Cyclophosphamide (CP) is used as an anticancer drug in chemotherapy. This drug has some side effects including anorexia, nausea, malfunction of sexual glands, azoospermia and oligospermia. The aim of this study was to evaluate the protective role of Heracleum persicum against gametogenesis damages in rats receiving cyclophosphamide. Thirty male Wistar rats (185-195 g) classified in five groups including control group (receiving 0.2 ml/day distilled water), second group (receiving 0.5 gr/kg/day intra-peritoneal cyclophosphamide), and three other groups (receiving same dose of cyclophosphamide intra-peritoneal with different doses of oral alcoholic extract of Heracleum persicum). The experiment lasted 21 days. At the end of experiment, the testes of rats were examined for histological and morphometric changes. The results showed that cyclophosphamide significantly reduced sperm counts, SI and TDI indices compared with the control group (P <0.01). However, groups who received the extract of Heracleum persicum in association with cyclophosphamide represented significantly improved parameters (P <0.01). Key words: Cyclophosphamide, Heracleum persicum, gametogenesis, rats. _______________________________________ http://pharmacologyonline.silae.it ISSN: 1827-8620 PhOL Dalouchi et al 198 (197-202) Introduction These drugs have many destructive side effects on Mammalian spermatogenesis is an ordered and healthy cells. Chemotherapy and radiation therapy well-defined process occurring in seminiferous are associated with changes in the male genital tract tubules of the testis. It is characterized by mitotical (5). Due to their intercalating with DNA causing spermatogonia divisions producing spermatocytes irreversible lesion, the drugs used in chemotherapy that proceed through meiosis to form a population contain highly toxic neoplastic agents that interfere of haploid cells (spermatids) over a period of with the cellular division (6). The high frequency of several weeks. Spermiogenesis is the haploid phase cellular divisions that occurs in the cells of the of spermatogenesis transforming spermatids into seminiferous epithelium makes the testis a highly spermatozoa (1). There are some disorders in vulnerable organ to the chemotherapeutical spermatogenesis located in the preservations or the drugs(6). Cis-diamminedichloroplatinum (II) or types of methods used for treatment (2). cisplatin (CIS) is a highly effective anti-neoplastic DNA Impairment in spermatogenesis as well as disorders alkylating agent used to treat many types of solid in the sperm production and function are among tumors, including testicular, ovarian, breast, lung, the most common causes of male infertility. Trauma bladder, head and neck. Furthermore, severe or anatomical defects in the reproductive system or testicular damage characterized by germ cell the use of certain drugs to treat disease lead to apoptosis, Leydig cell dysfunction and testicular impaired sperm production and ultimately to steroidogenic disorder are effectively cured by this impotency in men. (2). Infertility is one of the major measure (6,7) The use of medicinal plants has health problems in human life. Furthermore, recently been proposed to reduce the unfavorable approximately 30% of infertilities are due to a male effects of various therapeutic procedures. In factor. Several conditions can interfere with addition, many researches have been done in this spermatogenesis and reduce sperm quality and regard. A wide range of different biological activities, production. Other factors such as drug treatment, including antioxidant, anti-bacterial, anti-coagulation, chemotherapy, toxins, air pollutions and insufficient vasodilation, anti-inflammatory and anti-cancer vitamins intake have also harmful effects on medicinal plants have been identified, some of which spermatogenesis and sperm normal production. reduce the adverse effects of chemotherapy. Several studies have reported that antioxidants and Identification of the effects of medicinal plants on vitamins A, B, C, and E contained in diets can tissue and organs is a developing research domain protect the DNA of sperms from free radicals and (3,8). thus increase blood testis barrier stability(3). For example, Citrus fruits, such as oranges, contain By definition, cancer is a disease in which cells are components that have antioxidant potential and this abnormally divided and multiplied and they form factor has a useful effect on spermatogenesis and tumors. Tumors are known as hard lumps or sperm parameters (3). It is known that Croton ulcerated lesions that appear on surface of skin and Cajucara prevents testicular damage in rats muscles. Both benign and malignant anatomically undergoing chemotherapy (6). Ginkgo Biloba (GB) masses in forms of ulcers or abscesses that arise has been used in traditional Chinese medicine for between the muscle and bone and hard about 5000 years. Due to its antioxidant features, obstructions that arise in the internal organs (4). this herbal drug has been widely used in many Serious disorders, including benign and malignant situations. Studies show that GB could be involved in tumors, stem from internal injuries, emotional reducing toxicity in rats experiencing chemotherapy trauma, invasion of pathogenic factors-such as heat, drugs such as CIS (7). Heracleum persicum is a plant cold, dampness, dryness, or the accumulation of of Umbelliferae family and it is scientifically named toxins- that are often due to improper digestion and as Heracleum persicum. It is a perennial plant of the poor elimination of metabolic wastes. Besides, parsley dark that grows in high mountain areas, general therapeutic measures, such as especially in Azerbaijan, Fars, Ramsar, Hamedan, chemotherapy, are used to remove obstruction and Tehran, Damavand and other regions. This plant has produce new fresh blood (4). Chemotherapy thick roots and complete cut leaves. Persian’s prevents the division and proliferation of cancer Heracleum persicum is "Angdan". However, some cells and causes damage to this process. people call it "Anjudan". Heracleum persicum has Chemotherapy drugs are transported through blood two types. One type is an aromatic one that is named to cancerous cells and all the organs_______________________________________. as Heracleum persicum and Golapar. http://pharmacologyonline.silae.it ISSN: 1827-8620 PhOL Dalouchi et al 199 (197-202) The second kind is a funky plant that is called ethanol extract of Heracleum persicum orally. anghoze, angozhd, fat fingers, infected Haltit and The third group) the second experimental group( black Haltit. The chemical compounds of Heracleum received daily 5mg/kg/day intra-peritoneal injections persicum contain oil and this oil is volatized in the of cyclophosphamide and 1gr/kg/day ethanol extract form of powder so that foods can be aromatized. of Heracleum persicum orally. The plant of Heracleum persicum has many The fourth group) the third experimental group( medicinal features; it can fortify the stomach and received daily 5mg/kg/day intra-peritoneal injections other digestive organs as well as repulsive of cyclophosphamide and 2gr/kg/day ethanol extract gastrointestinal bloating. Furthermore, it can of Heracleum persicum orally. increase the gastric secretion and excretion of The fifth group) the fourth experimental groups( toxins in the body and milk of women. Finally, it is a received 5mg/kg/day intra-peritoneal injections of strong microbicide, disinfectant, and diuretic cyclophosphamide. material. Small amounts of these plants may have Twenty one days after the treatment, rats were effective medicinal effects on the reproductive anesthetized by chloroform and their abdomens system (8,9). were opened and the testes were placed in 10% Having regarded the traditional medicine, the formalin for histological supplying. Having prepared present study aimed to investigate the effect of the tissue samples, the microtome sections were Heracleum persicum in preventing adverse effects measured and the diameter of each section was of chemotherapy on spermatogenesis in rats. determined as 5 microns. Then, stained tissue slides were prepared with hematoxylin and Eosin. The Materials and Methods prepared slides were examined via using an optical Animals microscope with different magnifications. Thirty adult male Wistar rats (weighing 185 to 195 g) were provided from Tabriz Medical University. All Statistical analysis the rats were healthy and not infected with virus or The parameters considered in this study were bacteria and the keeping place of them had natural spermatid, sperm construction index (SI) and tubular light and darkness. They were sufficiently provided differentiation index (TDI) respectively. Data analysis with food and water except in experimentation was performed using software SPSS.19. Furthermore, time. Duncan's method was used in order to compare the means. Analysis of variance date showed that there Chemicals were significant differences among the studied